Akpachara anya maka polyacrylamide gel electrophoresis

· polyacrylamidejel kwesịrị site na acrylamide monomer, polymerization mmalite ihe, ihe na-akpali akpali, na ikike nke nnu na ngwakọta ngwakọta.

· acrylamidena BIS (N, N '- methylene double acrylamide) bụ monomer ụdị gel matrix.

· ammonium persulfate na-amalite nrapado polymerization usoro. Nhazi nke gluu chọrọ 10% ammonium persulfate ngwọta kwadebere na mmiri. Ọtụtụ n'ime ozi ahụ gosipụtara mkpa maka iji arụ ọrụ eme ihe. Otú ọ dị, 10% ngwọta nwere ike tinye na 4 ℃ ruo ọtụtụ izu na-enweghị ịrịba ọnwụ nke ọrụ. Mee ihe ruru 10 ml ma tụfuo mgbe gluu na-adakọtaghị.

Ndụmọdụ: Pasent nkeacrylamidena usoro gluu na protein gluu abụghị otu. Ọ bụrụ na ị na-eji acrylamide premade: ngwọta BIS, gbaa mbọ nweta karama kwesịrị ekwesị.

· TEMED (N, N, N', N'- tetramethyl ethylenediamine) bụ ihe na-akpali akpali, na karama agba aja aja, na-etinye n'ime friji. Tinye obere oge tupu ịwụsa gluu.

· polyacrylamide electrophoresis eji iko na electrophoresis kwesịrị ịsacha tupu na mgbe oge ọ bụla. Mgbe electrophoresis gachara, jiri ahịhịa dị nro na akwa sachaa n'ime mmiri ncha dị ọkụ, kpochaa ya na mmiri distilled wee guzo ka ọ kpọọ nkụ.

· mmiri na uzuzu nwere ike ịkpata na oghere polima. Tupu electrophoresis, hichaa efere iko ahụ na ihe nhicha iko ma hichaa ya na ahịhịa dị nro. Na-asa mmiri na-ekpo ọkụ ma kpochaa nke ọma na mpempe akwụkwọ. Ichacha na 70% ethanol tupu ihichapụ akwụkwọ na-enyere aka ihicha na ngwa ngwa ihicha. Tinye ihe atụ nke acrylamide n'otu n'otu: BIS, mmiri, ngwọta nchekwa, ammonium persulfate, TEMED. Ghichaa nke ọma ma wụsa ozugbo.

Ọ dịghị mkpa ịkwanye polyacrylamide tupu polymerization. (A na-etinye Acrylamide n'ime oghere iji wepụ ihe nfụkasị n'ihi na oxygen na-egbochi polymerization.)

Atụmatụ ntụle ntụ ntụ kwụ ọtọ.

· tinye mpempe oji n'ime igbe dị n'okpuru, ndabere ojii iji mee oghere nlele iji hụ nke ọma.
Gluo tank jupụtara ebe nchekwa, dị nnọọ n'elu colloid.
· ọ bụrụ na nsọtụ nwere ọkụ, gbanye ọkụ, ka ìhè na-enwu colloid. Denye ihe atụ n'ime pipette.
· iji ngwaọrụ ọkpọkọ akpaka.
· na 10-200 mu 1 mmiri mmiri na-akpụ akpụ nwere ike iji mee ihe n'ọtụtụ n'ime ihe ndị dị na sample. Maka obere oghere nlele (ihe na-erughị 10μ1), isi pipette ogologo ejiri maka usoro gluu na-adaba adaba.

· pipetting dị nnọọ mikpuru n'ime sample, na-ekuku ume nwayọọ nwayọọ nwayọọ mmiri mmiri. Ihe nlele ahụ nwere ike ịpụta na ọ na-arapara na glycerin, na ịgbapụta ngwa ngwa nwere ike ịdọta afụ ikuku n'ime isi pipette.

· samples mgbe ikuru ọkpọkọ isi, ga-akpali mmiri mmiri isi nwayọọ na nsọtụ nke ọkpọkọ ma ọ bụ na-ehichapụ akwụkwọ na-aṅụ isi mmiri mmiri n'èzí nke ụmụ irighiri mmiri. Kpachara anya ka ị ghara iri ihe nlele ahụ.

Tinye ihe nlele n'ime oghere nlele

· ngwaọrụ pipetting na-enwe ntakịrị nrụgide, mee ka ihe nlele ahụ na-ebugharị isi mmiri na-ejubiga ókè.
· isi pipetting etinye n'ime ihe nchekwa ahụ, dị ntakịrị elu karịa oghere ntụpọ, na-echekwa nrụgide dị mma. Enwere ike itinye ọnụ nke pipette n'ime obere oghere.
· ji nwayọọ nwayọọ na nwayọọ na samples pụta. A na-etinye ọnụ ọnụ pipette n'elu oghere ihe nlele, ihe nlele ahụ ga-abanyekwa n'ime oghere ahụ. Ka ihe nlele ahụ mikpuo ka o mejuo oghere nlele kama ịkwanye ya.
· otu ugboro ikpeazụ dobe sample na isi mmiri mmiri, mmiri mmiri ga-aga n'ụkwụ nke abụọ, jiri nwayọọ nwayọọ bulie mmiri mmiri na-aga, pụọ na nchekwa.

Kedu ka esi eme nlele gluu kwụ ọtọ?

· vetikal gluu ebe sample oghere kpụrụ n'etiti abụọ iberibe iko. N'ime gluu dị ezigbo mkpa, enweghị ike itinye isi pipette n'etiti efere iko abụọ. Lelee glycerin! Tinye isi pipette n'elu oghere nlele na ihe nlele ahụ ga-adaba n'ime oghere ahụ.
· atụ sample tupu, jide n'aka na-etinye vetikal nke polypropylene acyl gel sample oghere na-ichacha ọcha. Wepu acrylamide na-enweghị polymerized na mmiri nke nwere ike ịpụta na ala nke oghere nlele ahụ. Mmiri ahụ nwere ike ime ka oghere nlele ahụ dị ntakịrị. Jiri sirinji 25ml ma ọ bụ 50ml na agịga nlele 18. Wunye n'ime ihe nchekwa electrophoresis ma jiri nlezianya kpochapụ oghere nlele ahụ.
· ọ nwere ike isi ike ịhụ oghere nlele, ma n'otu aka ahụ, ndị ọzọ dị mfe. Ọ bụrụ na enwere oghere dị oke, enwere ike ịnwale ihe nchekwa ihe na bromophenol blue.